US 11,725,200 B2
Method for isolating RNA with high yield
Martin Schlumpberger, Hilden (DE); and Gabriele Christoffel, Hilden (DE)
Assigned to QIAGEN GMBH, Hilden (DE)
Appl. No. 16/650,598
Filed by QIAGEN GmbH, Hilden (DE)
PCT Filed Sep. 27, 2017, PCT No. PCT/EP2017/074490
§ 371(c)(1), (2) Date Mar. 25, 2020,
PCT Pub. No. WO2019/063071, PCT Pub. Date Apr. 4, 2019.
Prior Publication US 2021/0102189 A1, Apr. 8, 2021
Int. Cl. C12N 15/10 (2006.01)
CPC C12N 15/1003 (2013.01) 19 Claims
 
1. A phenol-free method for isolating a nucleic acid from a sample, said method comprising the following steps:
a) adding a precipitation buffer to a sample to prepare an acidic precipitation mixture wherein said precipitation buffer comprises a metal cation precipitant and a buffering agent, has a pH value of 4.0 or less and does not comprise an organic solvent selected from aprotic polar solvents and protic solvents and wherein the acidic precipitation mixture comprises the metal cation precipitant in a concentration of less than 200 mM and precipitating proteins;
b) separating the precipitate from the supernatant, wherein the supernatant comprises small RNA having a length of less than 200 nt and large RNA having a length of at least 1000 nt; and
c) isolating a nucleic acid from the supernatant.